By Vladimir V. Shuvaev, Thomas Dziubla, Rainer Wiewrodt, Vladimir R. Muzykantov (auth.), Christof M. Niemeyer (eds.)
Specialist laboratorians replace the vintage bioconjugation equipment and introduce helpful new options that transcend natural biomedical functions to incorporate components from complex natural synthesis, molecular biology, and fabrics technological know-how. those with ease reproducible tools conceal the practise of protein conjugates utilizing covalent and noncovalent conjugation, the synthesis of nucleic acid conjugates utilizing a number of labeling concepts, and ways to semisynthetic conjugates of proteins. extra chapters tackle the biofunctionalization of inorganic surfaces, together with the on-chip synthesis of peptide nucleic acids to generate microarrays for the high-throughput research of RNA and DNA, gold nanaoparticles, and carbon nanotube probes for atomic strength microscopy.
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Additional info for Bioconjugation Protocols: Strategies and Methods
Note that electrospray ionization mass spectrometry is not easily applicable for the characterization of PEG–protein adducts. In fact, the polydispersivity of PEG makes the data interpretation rather complicated. PEG–protein titration, amino acid analysis, and NH2 titration by TNBS or fluorescamine allow quantification of the overall number of PEG chains attached to the protein in the sample. SDS electrophoresis and MALDI mass spectroscopy visualize each PEGylated species in the sample even if they still fail to differentiate among the different positional isomers.
The hydroxyl groups were reacted with divinylsulfone in alkaline conditions. The divinylsulfone was added at 10-fold molar excess to the hydroxyl groups in the polymer in 30 mL of anhydrous dichloromethane, with a total polymer weight of 5 g. 03 g of potassium tert-butyloxide. The reaction was conducted at room temperature for 16 h under a nitrogen atmosphere. The derivatized PDEAAm was isolated by precipitation in diethyl ether (see Note 2). 2. Streptavidin Site-Directed Mutagenesis 1. A double lysine E51K/N118K streptavidin mutant was constructed by a combination of site-directed cassette mutagenesis and polymerase chain reaction (PCR) mutagenesis.
After 1 h, animals were sacrificed and blood and organs extracted and analyzed for radioactivity. A, 125I-anti-ICAM (black bars) or anti-ICAM/125I-tPA (hatched bars), but not free 125I-tPA (white bars) accumulate in the lung, liver, and spleen after intravenous injection. B, Comparison of biodistribution of anti-ICAM/125I-tPA after injections via the tail vein (hatched bars) or the left ventricle (black bars). Data are presented as mean ± SD, n = 4–9 animals per determination. Internalization and Pulmonary Targeting 33 Fig.
Bioconjugation Protocols: Strategies and Methods by Vladimir V. Shuvaev, Thomas Dziubla, Rainer Wiewrodt, Vladimir R. Muzykantov (auth.), Christof M. Niemeyer (eds.)